Molecular Combinations for Cancer or Other Disease Treatment

ABSTRACT

A method and medicine for treating leukemia or other cancer or disease condition including administering periodic doses of a combination of one or more of emodin, rhein or rhapontin with one or more of aucubin, digoxin or beta-sitosterol, and/or one or more of carvacrol, vanillic acid or sitosterol.

PRIORITY

This application is a Division of U.S. patent application Ser. No.13/218,437, filed Aug. 25, 2011; which is a Continuation of U.S. patentapplication Ser. No. 13/152,039, filed Jun. 2, 2011; which is aContinuation-in-Part of U.S. patent application Ser. No. 13/018,435,filed Feb. 1, 2011, now U.S. Pat. No. 8,541,382; which claims thebenefit of priority to U.S. provisional patent application No.61/413,430, filed Nov. 13, 2010.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates generally to disease treatments, includingtreatments for cancer including skin cancer and leukemia, as well as forautoimmune diseases such as psoriasis, chrone, ulcerative colitis,arthritis and diabetes. More specifically it concerns the use ofcombinations of certain herbs, certain herbal extracts and/or certainmolecular components of certain herbs to reduce white cell count.Example compounds and/or molecules include cardiac glycosides such asDigoxin and Quabain in combination with Emodin and its analogs for thetreatment of disease. Example herbs that may be combined and/or fromwhich certain compounds and/or molecules may be extracted and combinedinclude Da Huang and Sheng Di Huang, as well as Jin Yin Hua, and Mu DanPi, Di Gu Pi, Xian He Cao, and Chun Gen Pi.

2. Description of the Related Art

Herbal medicines are still prevalent, and serve the medicinal needs of alarge population around the world. The global herbal medicine market iscurrently worth around $30 billion. There is an increased effort for theisolation of bioactive phytochemicals from herbs for their possibleusefulness in the control of various ailments. Determining molecularstructure and mechanisms of action of bioactive phytochemicals areequally important for providing the evidence for their efficacy as wellas herbal preparations, which could also potentially lead to thepharmaceutical development of synthetic or semi-synthetic drugs. Aboutherbal medicines in cancer studies, several previous studies demonstratethat certain phytochemicals present in medicinal herbs exert anti-canceractivities. Among three structurally related anthraquinones, includingemodin, physcion, and chrysophanol, emodin showed the most potentcytotoxic effects on tumor cells. Emodin(1,3,8-trihydroxy-6-methylanthraquinone) and aloe-emodin,1,8-dihydroxy-3-hydroxymethyl-9,10-anthracenedione as shown achemotherapeutic activity in vitro and in animal models for thetreatment of various types of cancers, including bladder cancer, lungcancer, and hepatoma, and leukemia. Aloe-emodin is a naturalanthraquinone compound that is present in some traditional medicinalplants such as Rhei Rhizoma and Rheum palmatum, Interestingly,aloe-emodin has been found to have lesser cytotoxicity towards thenormal human cells.

Plant-derived cardiac glycosides such as digoxin are used for thetreatment of congestive heart failure and other cardiac disorders. Theirmain pharmacologic actions are mediated through inhibition of the sodiumpump, Na+- and K+-dependent ATPase (NKA) 1 (EC 3.6.3.9). NKA, aubiquitous membrane cationic transporter protein, controls normalmembrane potential in all eukaryotic cells by maintaining high K+ andlow Na+concentrations. It consists of a catalytic subunit and a βglycoprotein subunit. Studies have suggested that plant-derived cardiacglycosides regulate some cellular processes, such as proliferation andapoptosis, in a variety of cancer cells.

The following are incorporated by reference: WO2004/052294A2,WO2007/130124A1, and US20090018088.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates the effect of on proliferation of digoxin, Emodin,and their combination on NB4 leukemic cells.

FIG. 2 illustrates the effect of on proliferation of digoxin, Emodin,and their combination on HL-60 leukemic cells.

FIG. 3 illustrates the effect of on proliferation of digoxin, Emodin,and their combination on Jurkat T cell leukemic cells.

FIG. 4 illustrates the effect of on proliferation of digoxin, Emodin,and their combination on HT29 colon cancer cells.

FIG. 5 illustrates the effect of proliferation of digoxin, Emodin, andtheir combination on PC3 prostate tumor cells.

FIG. 6 illustrates the effect of proliferation of Digoxin, Emodin andtheir combination on MDA 435 Breast Cancer cells.

FIG. 7 illustrates the effect of proliferation of Digoxin, Emodin andtheir combination on MDA 435 cell lines of Melanoma and Breast Cancer.

FIG. 8 illustrates the effect of proliferation of Digoxin, Emodin andtheir combination on Heparanase activity.

FIG. 9 illustrates the effect of proliferation of Digoxin, Emodin andtheir combination on HMVEC activity.

FIGS. 10-11 illustrates the effect of proliferation of Digoxin, Emodinand their combination on NB-4 Cell Line for AML (Acute Milo Leukemia).

FIG. 12 illustrates the effect of proliferation of Digoxin, Emodin andtheir combination on HL-60 Cell Line.

FIG. 13 illustrates the effect of proliferation of Digoxin, Emodin andtheir combination on Jurkat cell line.

FIG. 14 illustrates the effect of proliferation of Digoxin, Emodin andtheir combination on HT-29 cell line.

FIG. 15 is a bar chart that illustrates the effects of certain herbs andcombinations of herbs on live cancer cells, including (C1) Sheng DiHuang—Rhemannia; as well as (C2) the combination of Jin YinHua—Lonicerae, Da Huang—Rhei, Mu Dan Pi—Moutan, and Di Gu Pi—CortexLycii, and (C3) the combination of Xian He Cao—Agrimoniae and Chun GenPI—Ailanthi.

FIGS. 16-18 show plots of growth of cancer cells versus dilution factorfor each of 18 different herbs.

FIG. 19 is a bar chart that illustrates the effects of certain herbs andcombinations of herbs on live cancer cells.

DETAILED DESCRIPTIONS OF THE EMBODIMENTS

A method of treating leukemia or other cancer is provided. The treatmentincludes administering to a patient diagnosed with the leukemia or othercancer a regimen that includes periodic doses of a combination of herbs,or herbal extracts, or compounds or molecules extracted from certainherbs or herbal extracts. Seven such herbs when used in certaincombinations exhibit enhanced potency in killing cancer cells and/orreducing white cell counts. These seven herbs include Da Huang, Sheng DiHuang, and Jin Yin Hua, as well as Mu Dan Pi, Di Gu Pi, Xian He Cao, andChun Gen Pi.

Administration in a treatment regimen of certain combinations of two ormore of these herbs serve to reduce white cell count, and are providedas embodiments herein. Specific embodiments include advantageouscombinations of Da Huang with Sheng Di Huang, as indicated below and inU.S. patent application Ser. No. 13/018,435, which is incorporated byreference, as well as the combination of Da Huang and/or Sheng Di Huangwith Jin Yin Hua. Further embodiments include combinations of Da Huangand/or Sheng Di Huang with one or more of Mu Dan Pi, Di Gu Pi, Xian HeCao, and/or Chun Gen Pi. Other embodiments include combinations of DaHuang and/or Sheng Di Huang with Jin Yin Hua and one or more of Mu DanPi, Di Gu Pi, Xian He Cao, and/or Chun Gen Pi. Further embodimentsinclude combinations of Jin Yin Hua with one or more of Mu Dan Pi, Di GuPi, Xian He Cao, and Chun Gen Pi. Other embodiments include combinationsof two of more of Mu Dan Pi, Di Gu Pi, Xian He Cao, and/or Chun Gen Pi.

Contained within each of the seven herbs are several molecularconstituents. An observed reduction of the white cell count owing to atreatment regimen of periodic doses of a combination of the seven herbscan be as a result of various combinations of active molecules, e.g.,emodin and digoxin which are contained in Da Huang and Sheng Di Huang,respectively. Advantageous effects of the regiment may be bolsteredfurther combinations with active ingredients and/or by one or morebuffer molecules or one or more molecules serving as some helpfulvehicle for the active molecules. The chemistries and pharmacologies ofthe seven herbs are summarized below.

In certain embodiments, certain parts of the seven herbs are used suchas the roots, stems, leaves, husks, branches, barks, sap, or kernels orcombinations thereof. For example, for Da Huang at least the root may beused. For Sheng Di Huang, at least dried root tuber may be used. For JinYin Hua, at least dried flower may be used. For Mu Dan Pi, at leastdried root bark may be used. For Di Gu Pi, at least dried root bark maybe used. For Xian He Cao, at least dried aerial part of Agrimonia PilosaLedeb may be used. For Chun Gen Pi, at least dried bark of the root orstem may be used.

In certain further embodiments, the two herbs Sheng Di Huang and DaHuang, or two or more of the three herbs including Jin Yin Hua, orcombinations of the seven herbs including Mu Dan Pi, Di Gu Pi, Xian HeCao and/or Chun Gen Pi with any one or more of the three herbs, may becombined with any one or more, even all in certain embodiments, of anadditional eleven herbs including Zi Cao, or radix arnebiae (arnebiaroot) or radix lithospermi (gromwell root), Xuan Shen, or radixscrophulariae (figwort root), Shi Gao or gypsum fibrosum (gypsum), BaiShao, or radix paeoniae alba (white peony root), Chi Shao or radixpaeoniae rubra (red peony root), Hong Hua or flos carthami (safflower),Da Qing Ye or folium isatidis (woad leaf), Qing Dai or indigo naturalis(natural indigo), Bai Zhu or rhizoma atractylodis macrocephalae(largehead atractylodes rhizome), and Shi Wei or folium pyrrosiae(shearer's pyrrosia leaf), Rou Gui or cortex cinnamomi (cinnamomumbark).

Da Huang Radix Et Rhizoma Rhei (Rhubarb) Chemistry

Da Huang contains free antraquinones, anthraquinone glycosides, andbianthrones. Among the free antraquinones contained within Da Huang arealizarin, aloe emodin, chrysophanol, citreorosein, emodin, laccaic acidD, physcion, and rhein. Among the anthraquinone glycosides containedwithin Da Huang are1,8-dihydroxy-3-methylanthraquinone-1-O-β-D-glucoside (Palmatin), aloeemodin 1′-O-β-D-glucopyranoside, aloe emodin 1-O-β-D-glucopyranoside,chrysophanol 1-O-β-D-glucopyranoside (chrysophanein), chrysophanol8-O-β-D-glucopyranoside, emodin 1-O-β-D-glucopyranoside, emodin3-O-β-D-glucopyranoside (Glucoemodin), emodin 8-O-β-D-glucopyranoside,physcion 1-O-β-D-glucopyranoside, physcion 8-O-β-D-gentiobioside,physcionin, and rhein 1-O-β-D-glucopyranoside. Among the bianthronescontained in Da Huang are aloe emodin bianthrone, chrysophanolbianthrone, palmidins A-C, rheidins A-C, sennidins A, B and C andsennosides A-F.

Da Huang also includes other compounds including2-(-2-hydroxy-propyl)-5-methyl-7-hydroxy-chromone,2-(-2-hydroxypropyl)-methyl-7-hydroxy-chromanone,2,5-dimethyl-7-hydroxychromone,2-methyl-5-carboxymethyl-7-hydroxychromone, 3 napthalenes,3,5,4′-trihydroxystilbene 4′-β-D-(2″-0-galloyl)-glucopyranoside,3,5,4′-trihydroxystilbene 4′-O-β-D-(6″-O-galloyl)-glucopyranoside,4′-O-methylpiceid, Rhapontin, Rheinosides A-D, Stilbene gakkates3,5,4′-rtihydroxystilbene 4′-O-β-D-glucopyranoside, Stilbene piceid andTannins.

Pharmcalogy Purgative Effect of Da Huang

Da Huang is well known as a purgative agent. The active constituents arethe combined anthraquinones, especially sennosides. The content ofsennosides correlates with the purgative activity of rhubarb. Table 1illustrates the oral purgative ED50 values of the anthraquinones.

TABLE 1 ED₅₀ Anthraquinones ED₅₀ (mg/kg) Anthraquinones (mg/kg)Sennoside A 13.5 Aloe-emodin-8-glycoside 71.6 Sennoside B 13.9 Emodinmonoglucoside 103.6 Sennoside C 13.3 Aloe-emodin 59.6 Sennoside D 13.8Rhein 97.5 Sennoside E 13.5 Emodin >500 Sennoside F 16.1 Physcion >500Rhein-8-glucoside 20.0 Chrydophanol >500

Studies on the mechanisms of action found that sennosides actpredominantly on the large intestine. The most potent purgative activitywas obtained from the rhubarb extract or Sennoside A by gastricaladministration and from sennidin by intravenous route. Inhibition of theintestinal flora in mice with chloramphenicol significantly decreasedthe activity prosthetic sugar group of the anthraquinone glycosidesprevented the anthrone from being oxidized before they are transportedinto the large intestine and hydrolysed by the bacterial enzyme intofree sennidins. It was found that sennosides are hydrolysed by microbialβ-glycosidase in a stepwise fashion to the corresponding sennidins via8-monoglycosides. The resulting metabolites sennidins were furtherreduced, possibly by a reductase bound to cell membranes of intestinalbacteria, to rheinanthrone as the purgative principle.

Ligation of the junction of the large and small intestines failed toprevent anthraquinone glycoside producing a purgative effect in thelarge intestine. Oral administration of rhubarb started to produceeffect 6-8 h later. These results suggest that there is also a largepart of anthraquinone glycosides absorbed in the small intestine andtransformed in the liver before they act on the pelvic plexus andproduce peristalsis and purgation.

On the other hand, small doses (0.05-0.3 g by oral administration)rhubarb caused constipation because of its high content of tannins. Theconstipation effect can be prevented by decocting rhubarb together withHuang Lian (Rhizoma Coptidis). This is because the tannins andberberine, the main constituent of Huang Lian, form gelatinousprecipitates during decoction.

Antimicrobial Effect of Da Huang

Tested by mixing virus with dilutions of aloe emodin for 15 min at 37°C., herpes simplex virus type 2 and type 3, varicella-zoster virus,pseudorabies virus, influenza virus were inactivated. Electronmicroscopic examination of the virus demonstrated that the envelopeswere partially disrupted, indicating that it is directly virucidal toenveloped viruses. Emodin and rhein showed antiviral activity againsthuman cytomegalovirus (HCMV) strain AD-169. When tested against aganciclovir-resistant strain of HCMV, the EC50 value for rhein wassuperior to the value obtained for the AD-169 strain. The aqueousextract of R. palmatum inhibited hepatitis B virus (HBV) polymeraseactivity and to bind hepatitis B virus surface antigen (HBsAg).Intravenous dose of 50 mg/kg of the extract to duck hepatitis B virus(DHBV) carrier ducklings showed antiviral activity against DHBV usingserum DHBV DNA level and DHBV DNA polymerase activity as antiviralindicators.

Rhubarb exhibited inhibition against staphylococci, Streptococcushemolyticus, Corynebacterium diphtheriae, Bacillus subtilis, B.brucellosis, B. mycoides, B. smegatis, Mycobacterium graminis, Yerseniapestis, Salmonella tophi, S. paratyphi, Shigella dysenteriae andNeisseria gonorrhea. Staphylococci and Neisseria gonorrhea were mostsensitive to the herb. The main antibacterial components were theanthraquinone derivatives with the structure of1,9-dihydroxyanthraquinone. 3-Carboxyrhein, hydroxyaloe-emodin andhydroxyemodin showed the most potent antibacterial activity. Thebacteriostatic concentrations of rhein, emodin and aloe-emodin againststaphylococci, streptomycin, Corynebacterium diphtheria, Bacillussubtilis, B. anthracis and Salmonella tophy were mitochondrialrespiratory chain of microorganisms. Respiration of Staphylococcusaureus was strongly inhibited by emodin, aloe-emodin and rhein. Rhein,emodin and rhein specifically interfered with the redox function NADHdehydrogenase.

The aqueous, ethanolic and ether extracs of rhubarb are also antifungalagainst many pathogenic fungi, including Achorion schoenleini,Trichopphyton concentricum, T. violaceum, T. gypsum, Nocardia asteroids,Epidermophyton flocosum and Sporotrichum schenckii. The decoction ofrhubarb exhibited inhibition against the influenza virus. The minimaleffective dose in chicken embryo in vitro and semi in vivo was 5 mg perembryo.

Antineoplastic and Antimutagenic Effects of Da Huang

Intraperitoneal administration of 75 mg/kg of emodin produced a 45%inhibition against the mammary carcinoma of mice. The inhibition ratesof 5 mg/kg of rhein and emodin against murine melanoma were 76% and 73%.Rhein, emodin and aloe-emodin inhibited murine leukemia P388 in vivo,increasing the survival time and decreasing the ascites volume. Theyalso inhibited Ehrlich ascites carcinoma, Emodin was a strong inhibitorof respiration in Ehrlich ascites carcinoma cells, with an ED50 of 20μm/ml. Cellular respiration in leukemia L1210 cells was also inhibited.Palmatin, Cysophanein and physcionin also exhibited moderate cytotoxicactivity against several types of carcinoma cells.

The extract of the herb from R. palmatum and emodin induced adose-dependent decrease in the mutagenicity of benzo(a)pyrene [B(a)P,],2-amino-3-methylimidazo(4,5-f) quinoline (IQ) and3-amino-1-methyl-5H-pyrido(4,3-b)indole (trp-P-2) in Salmonellatyphimurium TA98. It was further found that emodin reduced mutagencityof IQ by direct inhibition of the hepatic microsomal activation and notby interaction with proximate metabolites of IQ and/or by modificationof DNA repair processes in the bacterial cell. Emodin also markedlydecreased the mutagenicity of 1-nitropyrene (1-NP) in a dose dependantmanner in Ames-microsomal test with S. typhimurium TA98 and thegenotoxicity in SOS chromotest with E. coli PQ37. Furthermore, emodinsignificantly inhibited the formation of 1-NP DNA adducts in S.typhimurium TA98. The results suggest that emodin acts as a blockingand/or suppressing agent to reduce the direct-acting mutagenicity of1-NP.

Hemostatic Effect of Da Huang

Rhubarb is also in TCM as a hemastatic agent. The hemastatic activityhas been proved experimentally and clinically. Rhubarb is effective forboth external and internal hemorrhage. It was effective in the treatmentand prevention of experimental gastric bleeding and ulcer formation inrats. Significant therapeutic effects of the powdered rhizome of R.palmatum in the treatment of gastrointestinal bleeding were alsoreported. It reduces coagulation time and the permeability and fragilityof capillaries. It increases fibrinogen and promotes bone marrow toproduce platelets.

Immunosuppressive Effect of Da Huang

Emodin at 3×10-7 −3×10-4 M dose-dependently suppressed the responses ofhuman mononuclear cells to phytohemagglutinin and mixed lymphocytereaction. It was further found that after exposure to emodin (10-6 M)the production of interleukin-1 (IL-1) and interleukin-2 (IL-2) and theexpression of IL-2 receptor were all decreased, Emodin may be a newtemplate for the development of better immunosuppressive agents for useagainst transplantation and autoimmune disease.

Choleretic Effect of Da Huang

Rhubarb can stimulate construction of the gallbladder and relax Oddi'ssphinctercan, thus promoting bile secretion. It also increases thecontents of bilirubin and bile acid.

Other Effects of Da Huang

Oral administration if emodin and rhein provoked marked diuretic,natriuretic and kaliuretic effects in rabbits. Oral administration ofrhubarb increased urinary excretion of sodium and potassium, alkalizingurine to a pH value as high as 8.4. Rhubarb also inhibits the activitesof pepsase, trypsase, pancreatic amylase and pancreatic lipase. Itlowers blood pressure and blood cholesterol levels. Rabbits with feverinduced by subcutaneous injection of the pneumococci responded withreduced temperature after oral administration of the decoction ofrhubarb.

Intraperitoneal administration of 15 mg/kg of emodin exhibitedantiinflammatory activity against carrageenin-induced pedal inflammationin rats. In the same dosage it also showed antiulcerative activityagainst pylorus-ligated, aspirin and immobilization stress-inducedgastric ulcer in rats. It decreased acid and pepsin output and augmentedmucus secretion in terms of total carbohydrate:protein ration in thegastric juice of aspirin treated pylorusligated rats, indicating thatthe antiulcerative effect of emodin may be due to this effect on gastricsecretion.

Mu Dan Pi Cortex Moutan (Paeony Bark) Chemistry

Mu Dan Pi contains Apiopaeonoside, Benzoyloxypaeoniflorin,Benzoylpaeoniflorin, Galloyloxypaeoniflorin, Galloypaeoniflorin,Mudanpiosides A-F, Oxypaeoniflorin, Paeconoside, Paeoniflorin,Paeonisuffral, Paeonisuffrone, Paeonol, Paeonolide, Pentagalloyglucose1,2,3,4,6, and Suffruticosides A-E.

Pharmacology Antimicrobial Activity of Mu Dan Pi

The decoction of the root bark exhibited a strong antibacterial activityin vitro against the following bacteria: Bacillus subtilis, Escherichiacoli, Salmonella typhi, S. paratyphi, Protues vulgaris, Pseudomonasaeruginosa. Staphylococcus aureus, Strephtococcus hemolyticus,Diplococcus pneumoniae and Vibrio cholerae. Paeonol was one of theantibacterial components; its MIC values were 1:2000 againstStaphylococcus aureus, 1:1500 against Bacillus subtilis and Escherichiacoli.

Anti-inflammatory Effect of Mu Dan Pi

The 70% methanolic extract of the root bark inhibited rat paw swellinginduced by carrageenin. Paeonol was found active in inhibiting rat pawswelling induced by dextran, acetic acid or carrageenin. It alsoinhibited the increase of intra-abdominal capillary permeability of miceand cutaneous capillary permeability of guinea pigs caused by aceticacid or 5-HT. On the other hand, the water soluble fraction free frompaeonol as well as the glycoside fraction also exhibited a significantinhibitory action on rat paw edema due to carrageenin. The water-solublefraction was also effective in either preventing or treatingadjuvant-induced arthritis in rats. The methanolic extract, theglycosidic fraction and paeonol inhibited blood platelet aggregation.ADP- or collagen-induced human plateet aggregation was inhibited bypaeonol. The formation of thromboxan B2 was also inhibited but theformation of 12-hydroxy-5,8,10,14-eocpsatetraenoic acid fromC-arachidonate was stimulated. Besides, paeonol inhibited the formationof prostanoids such as prostaglandins and thromboxanes fromC-arachidonate in rat peritoneal macrophages. Thus the anti-inflammatoryaction of the root bark was related to the inhibitory effects of paeonolon prostanoid synthesis.

Hypotensive Effect of Mu Dan Pi

The blood pressure of dogs with essential or renal hypertension wassignificantly reduced after oral administration of 5 g/kg of thedecoction of the root bark for 5 days and 10 g/kg for two more days. Theblood pressure of dogs with renal hypertension was also lowered afteroral of 10 g/kg of the decoction free paeonol for 10 days. Oraladministration of 0.5-1.0 g/kg of paeonol also produced hypotensiveaction renal hypertensive dogs and rats.

Effect on the CNS of Mu Dan Pi

Intraperitoneal or oral administration of paeonol decreased thespontaneous activity of mice, antagonized caffeine-induced hyperactivityand prolonged cyclobarital-induced sleep. At higher doses, paeonolcaused disappearance of the righting reflex in mice. It also antagonizedconvulsions due to cardiazol, strychnine, nicotine and electric shock.Furthermore, paeonol was found to have antipyretic and analgesicactivities. Paeonol decreased the body temperature of normal mice andthe mice with typhoid and paratyphoid vaccine-induced fever. Oraladministration of paeonol produced an analgesic effect against aceticacid-caused writhing and tail pain by pressing in mice.

Effect on Obesity of Mu Dan Pi

The aqueous extract of the herb was given as drinking water at theconcentration of 0.5% to (SLN×C3H/He) F1 obese mice between 3 and 32weeks of age. The treatment resulted in a significant decline,particularly in males, in food intake and in the Lee index. An index ofobesity, and furthermore an increase in glucose tolerance. Nosignificant difference was observed between the experimental and thecontrol groups in the serum free fatty acid levels. There was littledifference between groups in the weights of heart, liver, lung, spleenand major endocrine organs in both sexes and in the pattern of oestrouscycles in females, There results indicate that the herb protects againstobesity, especially in males, at least partly by a decrease in foodintake and an increase in glucose metabolism.

Other Effects of Mu Dan Pi

Paeonol exhibited anticholinergic and antihistaminic actions on isolatedileum of mice and guinea pigs. It also prevented stress ulcer in miceand inhibited gastric secretion in rats and spontaneous motility of ratuterus in situ. The extract of the root bark and paeonol were also ofantimutagenic activity. They decreased the frequency of mutationsinduced by 4-nitroquinoline 1-oxide in Escherichia coli WP2s.

Sheng Di Huang Radix Rehmanniae (Chinese Foxglove Root) Chemistry

Sheng Di Huang contains4-(α-L-rhamnopyranosyloxy)-3-methoxybenzoylajugol, Aceutoside, Ajugol,Aucubin, Campesterol, Castanosides A and F, Catalpol, Echinacoside,E-feruloylajugol, Isoacetoside, Jioglutoside A and B, Jionosides A&B,Leonuride, Mannitol, Melittoside, p-courmaroylagujol,p-hydorxybenzoylajugol, Purpureaside C, Rehmaglutins A, RehmaionosidesA-C, Rehmanniosides A-D, Rehmapicroside, Vanilloylajugol,Z-feruloylajugol, and β-sitosterol.

Pharmacology Effects on Adrenocortical Function and Cortisol Metabolismof Sheng Di Huang

The herb was able to stop the decrease of plasma corticosteroneconcentration due to administration of dexamethasone and prevent theadrenal cortex from atrophy. The corticosterone level in rabbitsreceiving dexamethasone was increased at week 4 and week 6 when the herbwas concurrently applied. No morphological changes were observed in thepituitary gland and adrenal cortex of rabbits receiving concurrenttreatment of dexamethasone and the herb. A single large dose (3 g/kg) ofthe root or together with two other herbs Zhi Mu (Rhizoma Anemarrhenae)and Licorice (0.9 g/kg each) given orally antagonized the inhibitoryeffect of dexamethasone on the pituitary-adrenal system of rabbits,thereby increasing plasma corticosterone level.

This mixture also antagonized the inhibitory action of dexamethason onthe early morning cortisol secretion peak in 12 normal subjects astested in diurnal dexamethason suppression test. The crude extract (8mg) of the root, when incubated with the liver sections of rabbits,protected cortisol from being reduced on the double bond between C4 andC5, and the ketone at C3 and being degraded of the hydroxyl groups atC17 and C21, and the ketone at C20, thus delaying the metabolism ofcortisol in the liver. When the herb was used simultaneously withexogenous adrenocortical hormones, plasma cortisol could still be keptat a nearly normal level. The mechanism is believed to be a kind ofcompetitive effect which influenced the binding of cortical hormone tothe receptors and affected the uptake of corticosteroid hormone by theliver cells, thereby slowing down the catabolism of cortisol.

Cardiovascular and Diuretic Effects of Sheng Di Huang

The effects of the herb on the heart were largely dependent on doses.There was no obvious cordial activity at 0.1 or 0.5% concentration. At1% concentration, cordial effect was observed in the isolated perfusedfrog heart. This action was more obvious in weak heart. Whenconcentrations were increased to 2-5%, the heart was inhibited. Itseffects on blood pressure was also dose-dependent. In an experiment withperfuesd vessels, 1-3% of the extract caused vasoconstriction while at5% vasodilation.

Intravenous injection of 2.5 ml of the root extract produced a diureticeffect in anesthetized dogs. This action may be related to the cordialand renal vasodilation activities.

Effect on Blood Glucose of Sheng Di Huang

Results in animal experiments have been inconsistent on the effect ofthe herb on blood glucose. Hypoglycemic effects of alcoholic extract ofthe root at a subcutaneous dose of 2 g/kg or an oral dose of 4 g/kg inrabbits were reported by early researchers. The result obtained from thesubcutaneous injection was more significant: the blood sugar wasdecreased to the lowest level 4 h after medication. Subcutaneousadministration of the alcoholic extract to rabbits also inhibited theprolonged hyperglycemic effect elicited by carbohydrates from the rootof Codonopsis pilosula (Dang Shen). Intramuscular administration of 20 gof the same extract also suppressed and prevented epinephrine-inducedhyperglycemia in rabbits. Other studies, however, reported that theaqueous or alcoholic extract could only reduce the blood glucose ofnormal rabbits and was not effective in hyperglycemia due toepinephrine. But there were also reports that the herb had no effect onthe normal blood glucose level of rabbits. The decoction or theethanolic extract at 6 g/kg had no effect on the normal blood glucosemeasurements of rabbits within 6 h of medication. Subcutaneousadministration of 20 g/kg of the same agents also failed to antagonizeepinephrine-induced hyperglycemia in rats. More recently, a weakhypoglycemic activity of rehmannioside D in spontaneous diabetic micewas reported.

Antiinflammatory and Immunosuppressive Effects of Sheng Di Huang

Formaldehyde-induced edema of rat paws subsided after oraladministration of the decoction or alcoholic extract at the daily doseof 10 g/kg for 5 days. However, another report claimed that only thedecoction and not the alcoholic extract had a significantanti-inflammatory activity. At the oral dose of 100 mg/kg, jionoside Band acetoside produced 36% and 18% suppression of hemolytic plaqueforming cells in the spleens of mice. In the same test conditions,intraperitoneal dose of 30 mg/kg of cyclophosphamide had a 52.5%suppression.

Hemostatic Effect of Sheng Di Huang

The coagulation time in rabbits was reduced after giving the yellowneedle crystal obtained from the ethanolic extract of the root.Intraperitoneal administration of 10 g/kg of the decoction or ethanolicextract, or oral administration of the charred herb shortened thebleeding time in mice with tail wounds.

Effect on Hemorheology of Sheng Di Huang

The effects of the herb on the hemorheology of inflammatory, thromosicand intact animals were examined. Oral administration of 200 mg/kg ofthe 50% ethanolic extract of the herb inhibited the reduction offibrinolytic activity erythrocyte deformability, the decrease inerythrocyte counts and the increase in connective tissue of the thoracicartery in a chronic inflammatory model, adjuvant-induced arthritis.However, it was ineffective on the development of edema in the arthriticrats and on acute and chronic inflammation. In addition, the extractinhibited the reduction of erythrocyte deformability but not thedecrease of coagulative factors in a thrombosic model, endotoxin-induceddisseminated intravascular coagulation (DIC). It also exhibited apromoting effect on erythrocyte deformability and fibrinolytic activityin intact rats. There results suggests that oral administration of theextract can prevent an inducement of impediment in the peripheralmicrocirculation of various chronic diseases through the improvement ofhemorheology.

Other Effects of Sheng Di Huang

Antiradiation, antifungal and antihepatotoxic activities have also beenobserved with extract of the root in animals. The 100% injectionsolution of the root given intraperitoneally at 1 ml daily for 6 daysmitigated platelet damage in rats caused by 600 rad of γ-irradiation.The aqueous extract of the root inhibited intro fungi mentagrophyton.Microsporum gypseum and M. audouini. The decoction of the root showedprotective effect in mice against CCI-caused liver intoxication. Oral orintraperitoneal administration of 10 g/kg of the decoction or thealcoholic extract potentiated the hypnotic effect of pentobarbitalsodium. Intraperitoneal dose of 20 g/kg of the decoction or thealcoholic extract protected mice from hypobaric hypoxia.

Jin Yin Hua Flos Lonicerae (Honeysuckle Flower) Chemistry

Jin Yin Hua contains 2,6,6-trimethyl-2-vinyl-5-hydroxytetrahydropyran,Benzyl-alcohol, Carvacrol, Cis andtrans-2-methyl-2-vinyl-5-(α-hydroxyisopropyl)-tetrahydrofuran,Epivogeloside, Eugenol, Geraniol, Hex-1-ene, Hex-3-en-1-ol,

Isochlogogenic acid b+c (two isomers of 3,4-dicaffeoyl quinic acid),

Isochlorogenic acid a (3,5-dicaffeoyl quinic acid), Linalool, Loganin,Lonicerin, Lonicerin, Luteolin, Methylcaffeate, Pinene, Saponins witholeanolic acid, Secologanin dimethylacetal, Secoxyloganin, Vanillicacid, Venoterpin (gentialutine),

Vogeloside, α-terpineol, and β-phenylethyl alcohol.

Pharmacology Antimicrobial Activities of Jin Yin Hua

The extracts of both the flower and vine inhibited in vitro thefollowing bacteria Staphylococcus aureus, Streptococcus hemolyticus,Escherichia coly, Shigella dysenteriae, Vibrio cholera, Salmonellatyphi, S. oaratyphi, Diplococcus pneumoniae, Neisseria meningitides,Pseudomonas aeruginosa and Mycobacterium tuberculosis. It alsopotentiated the action of penicillin against the drug-resistantStaphylococcus aureus. Chlorogenic acid and isochlorogenic acid arebelieved to be the chief antibacterial components of the flower.Luteolin also showed an antibacterial activity. More than half of themice receiving the LD dose of Pseudomonas aeruginosa or its endotoxinsurvived after given 7.5 g/kg of the injection solution of the flower byintraperitoneal administration. Intravenous administration of 6 g/kg ofthe distillate of the flower was also therapeutically effective inrabbits poisoned by the endotoxin of Pseudomonas aeruginosa.

Antifunal activity was observed with the aqueous extract of the floweragainst Microsporum ferrugineum and Nocardia asteroids. In the monolayerprimary culture of the epithelial cells of human embryonic kidney, thedecoction of the flower inhibited influenza virus, ECHO virus and herpesvirus.

Anti-inflammatory Effect of Jin Yin Hua

Intraperitoneal administration of 0.25 g/kg of the flower inhibitedcarrageenin-induced paw edema in rats. Given twice a day at 8 g/kg for 6days by Intraperitoneal injection, the extract of the flower showedantiexudative and antihyperplastic effects on croton oil-inducedgranuloma. Intraperitoneal administration of the injection solutionincreased the phagocytic activity of the inflammatory cells in mice. Thedecoction diluted to 1:1280 concentration was still able to promoteleukocytic phagocytosis.

Central Stimulant Effect of Jin Yin Hua

Oral administration of chlorogenic acid produced central stimulation inmice and rats in experiments using electric shock and revolving cage;the potency of the central stimulation was ⅙ that of caffeine. Noaddictive or synergistic action was observed when they were usedtogether.

Antilipemic Effect of Jin Yin Hua

Oral administration of 2.5 g/kg of the flower reduced the intestinalabsorption of cholesterol and the plasma cholesterol level. In vitroexperiments showed that the flower conjugated with cholesterol.

Other Effects of Jin Yin Hua

Intraperitoneal administration of an aqueous-ethanolic extract of theflower of L. japonica to mice on day 8 after mating decreased pregnancyin the test animals dose-dependently. Intrauterine and intra-amnioticadministration of the extract killed the fetuses in dogs and causedabortion in monkeys, respectively. The extract of the flower exhibited amild prophylactic effect against experimental gastric ulcer in rats whengiven orally. Large oral doses of chlorogenic acid increasedgastrointestinal peristalsis and promoted gastric and bile secretion.Chlorogenic acid had a stimulant effect on the isolated rat uterus.

Di Gu Pi Cortex Lycii Radicis (Wolfberry Bark) Chemistry

Di Gu Pi contains 5α-stigmastan-3 6-dione, Betaine, Cinnamic acid,Kukoamine A, Linoleic acid, Lyciumamide, Melissic acid, Sugiol, andβ-sitosterol.

Pharmacology Antipyretic Effect of Di Gu Pi

The aqueous or alcoholic extract of the herb, given orally or byinjection, produced a significant antipyretic effect in rabbits withfever induced by pyrogen. Betaine was also active. A strong antipyreticeffect was also exhibited by the aqueous fraction of the alcoholicextract at doses ranging from 0.75 to 7.5 g/kg equivalent of the crudedrug. The precipitates of the extract from lead salt also showedcomparable antipyretic activity to synthetic antipyretic analgesics.

Hypoglycemic Effect of Di Gu Pi

Oral administration of the decoction of the herb decreased blood glucoselevel in rabbits by 14% in average; this action lasted for 7-8 h. Thepeak action was observed 3 to 4 h after administration. It was also lesseffective when give subcutaneously. Subcutaneous dose of 6 g/kg of theextract elicited a mean reduction of 14% of the blood glucose of rabbitsafter 1 h.

Hypotensive and Anticholesterolemic Effect of Di Gu Pi

The decoction, macerate, tincture and injection solution of the herbproduced a significant hypotensive effect in anesthetized dogs, cats,rabbits by intravenous or intramuscular administration and inanaesthetized rats by oral administration. Repeated intravenousadministration at lower doses did not induce rapid tolerance.Intravenous injection of 0.375 g/kg of the injection solution resultedin sudden drop of blood pressure and death of anesthetized dogs.Bradycardia, prolongation of PR interval and depressed T wave in the ECGwere observed. Kukoamine A induced hypotension in rats when givenintravenously at a dose of 5 mg/kg.

Daily oral administration of 10 g/kg of the extract of the herb for 3weeks decreased the serum cholesterol in rabbits by 36.9% with littleeffect on triglyceride.

Antimicrobial Activity of Di Gu Pi

In the sensitivity test using the paper disc method, the decoction ofthe herb strongly inhibited Bacillus typhosus, Salmonella paratyphi A,and Shigella flexneri but was inactive against Staphylococcus aureus. Itwas a weak bacteriostatic against Mycobacterium tuberculosis. In theprimary monolayer tissue culture of the embryonic renal cells, thedecoction prevented the pathogenic changes in the cells due to Asianinfluenza virus A JK strain.

Effect on the Uterus of Di Gu Pi

The 100% injection solution of the bark showed stimulation effects onnormal rat uterus and isolated mouse uterus. The activity of 1 ml of thesolution was comparable to that of 0.054 unit of pituitrin.

Xian He Cao Herba Agrimoniae (Hairyvein Agrimonia Herb) Chemistry

Xian He Cao contains Agrimols A, B, C, D and E, Agrimoniin,

Agrimonolide, Agrimorphol, Apigenin-7-glucoside, Caffeic acid, Ellagicacid, Gallic acid, Luteolin-7-glucoside, Pendinculagin, Potentillin, andQuercetin

Pharmarcology Teniacidal Effect of Xian He Cao

The winter sprout of the herb is used in folk medicine to expel teniaand the active principle was found to be agrimophol. Agrimophol actsdirectly on the parasite. It inhibits the glycogenolysis, aerobic andanaerobic metabolism in the parasite.

The herb and agrimophol are also lethal to some other parasites, such asTrichomonas vaginalis, blood fluke and roundworm.

Antibacterial Activities of Xian He Cao

Six compounds, luteolin-7-glucoside, apigenin-7glucoside. Quercetin,ellagic acid, caffeic acid and gallic acid, isolated from the herb wereactive against bacillary dysentery. Combination use ofluteolin-7-glucoside and ellagic acid, apigenin-7-glucoside andapigenin-7-glucoside was more effective than the respective individualcompounds.

Antitumor Effect of Xian He Cao

Agrimoniin had antitumour activity when given as a pre- orposttreatment. A single dose of 10-30 mg/kg prolonged the life span ofmice bearing MM2 tumors or yielded cures when given intravenously ororally prior to or after tumor inoculation. Agrimoniin also inhibitedthe growth of MH-134 and Meth-A solid tumors in mice. It was stronglycytotoxic to MM2 cells in vitro, but the activity was almost completelyabolished by the addition of fetal calf serum to the culture.Intraperitoneal administration of agrimoniin increased the number ofperipheral white blood cells and the proportion of monocytes. Theantitumor activity of agrimoniin appeared to be due to its enhancementof the immune response.

Cardiovascular Effect of Xian He Cao

Intravenous administration of the alcoholic extract of the herbincreased blood pressure and stimulated respiration in anesthetizedrabbits and dogs, but the alcohol-soluble fraction of the aqueousextract lowered blood pressure in rabbits. Perfused into the bloodvessels of rabbit ear and frog hind limb, it caused vasoconstriction atlow concentrations and vasodilation at high concentrations. The extractand agrimoniin increased the heart rate and cardiac contractility offrogs and toads. On the other hand, the alcohol-soluble fraction of theaqueous extract inhibited the isolated frog heart.

Chun Gen Pi Cortex Ailanthi (Tree-of-Heaven Bark) Chemistry

Chun Gen Pi contains 1-(1′,2′-dihydroxyethyl)-4-methoxy-β-caboline,1-(2′-hydroxyethyl)-4-methoxy-β-carboline,1-(2-hydroxy-1-methoxy)-ethyl-4-methoxy-β-carboline,13(21)-dehydro-glaucarubinone, 13(21)-dehydroglaucarubolone,1-acetyl-4-methoxy-β-carboline, 1-carbamoyl-β-carboline,1-carbomethoxy-β-carboine, 1-hydroxycanthin-6-one,1-methoxycanthin-6-one, 1-methoxy-canthin-6-one-3-oxide,5-hydroxymethylcanthin-6-one, 6-methoxy-β-carboline-1-carboxylic methylester, Ailanthone, Ailantinols A and B, Amarolide, Amarolide 11-acetate,Canthin-6-one, Canthin-6-one-3-oxide, Chaparrinone, Chaparrolide,Glaucarubinone Quassinoids Δ13(18)-dehydro-glaucarubinone,Δ13(18)-dehydroglaucarubolone, Shinjudilactone, Shinjulactones A-N, andβ-carboline-1-propionic acid.

Pharmacology

Glaucarubinone and ailanthone showed amebicidal activity in vitroagainst the parasite Entamoeba histolytica. Some quassinoids markedlyinhibited the growth of chloroquine-resistant Plamodium falciparum.Glaucarubinone produced complete inhibition at 0.0006 μg/ml.

Ailanthone also showed potent antiulcer activity.

FIG. 15 illustrates the effect on live cells of three combinations ofherbs each in three concentrations 1:40, 1:20 and 1:10. C1 includesSheng Di Huang, C2 includes Jin Yin Hua, Da Huang, Mu Dan Pi and Di GuPi, and C3 includes Xian He Cao and Chun Gen Pi. FIG. 15 illustratesthat each of these three example combinations provides a reduction ofthe live cell numbers particularly in higher concentrations, while C1and C2 appear to be more effective than C3. Each of C1 and C2 includesone or two of the three herbs Sheng Di Huang, Jin Yin Hua and Da Huang,while C3 does not. In addition, the combination of four herbs in C2appears to be more effective than the single herb in C1.

FIGS. 16-18 illustrate plots for each of the eighteen herbs of growth asa percentage of control versus dilution factor. FIGS. 16-18 illustratethat the three herbs Sheng Di Huang, Da Huang and Jin Yin Hua are mosteffective, while the four herbs Mu Dan Pi, Di Gu Pi, Xian He Cao andChun Gen Pi are effective, and the eleven herbs Zi Cao, Xuan Shen, ShiGao, Bai Shao, Chi Shao, Hong Hua, Da Qing Ye, Qing Dai, Bai Zhu, ShiWei and Rou Gui are somewhat less effective.

FIG. 19 illustrates the effect on live cells of five combinations ofherbs each in two concentrations 1:40 and 1:80. Cal includes acombination of all eighteen herbs, C2 includes a combination of Jin YinHua, Da Huang, Mu Dan Pi and Di Gu Pi, C3 includes a combination of DaHuang, Sheng Di Huang and Jin Yin Hua, C4 and C5 include combinations ofthe other eleven herbs (Zi Cao, Xuan Shen, Shi Gao, Bai Shao, Chi Shao,Hong Hua, Da Qing Ye, Qing Dai, Bai Zhu, Shi Wei and Rou Gui) of theeighteen herbs, i.e., not in combination with any of the seven herbs(Sheng Di Huang, Da Huang, Jin Yin Hua, Mu Dan Pi, Di Gu Pi, Xian He Caoand Chun Gen Pi). Specifically, C4 includes Shi Gao, Shi Wei, Bai Zhu,Rou Gui, Hong Hua and C5 includes Zi Cao, Xuan Shen, Chi Shao, Bai Shao,Da Qin Ye, and Qing Dai. FIG. 19 illustrates that combinations of alleighteen herbs, as well as combinations of the three herbs (Sheng DiHuang, Da Huang and Jin Yin Hua) are most effective, while thecombination C2 was less effective, and the combinations C4 and C5 werestill less effective.

Herbal Ingredients Aloe Emodin

Aloe emodin has a molecular weight around 270.24 g/mol. As to itsanti-cancer activity, aloe emodin exhibits cytotoxicity in SCC oftongue, cervix cancer cells; and apoptoticity through MAPK-JNK cascadein hepatoma cells. Aloe emodin also tends to induce P53 and apoptosis.In addition, the cytotoxicity of aloe emodin induces effects inmelanoma, and gastric carcinoma. As to its anti-inflammatory activity,aloe emodin exhibits anti TNF and anti virality in envelopedviruses—HSV, PSV, VSV, and INF. Aloe emodin also decreases COX2 and INOSexpression in inflammation, and increses IFN in JEV and EV71. Aloeemodin may also be ingested for its properties as a laxative. Use ofaloe emodin can induce nausia, and intestinal contraction causingabdominal pain. Long use of anthraquinones can lead to kidney and liverdamage. The half-life of aloe emodin is about 78 min.

Chrysophanol

Chrysophanol has a molecular weight around 254.24 g/mol. As to it santi-cancer activity, chrysophanol exhibits necrosis in hepatic cancercells, including ATP change and ROS cascade. Chrysophanic acid inhibitsEGFR in colon cancer cells. As to its anti-inflammatory activity,chrysophanol can serve as an antiseptic, bactericide, candidicide,and/or cathartic. In addition, chrysophanol can be used as an anti-staphaurus and an anti-bacilus subtilis. Chrysophanol can be used to suppressthe activation of NF-kB. and caspase-1 in LPS-stimulated macrophages.Chrysophanol is also an anti polio virus compound. Chrysophanol an beused as a hemostat and as a pesticide, and can be used in the treatmentof menorrhagia, including bleeding following abortion, epistaxis,functional uterine bleeding and thrombocytopenia. Chrysophanol acts as apurgative, and can be used to converts aloe emodin through P450 in theliver. Emodin and chrysophanol may be ingested in combination as an anticancer treatment agent. Chrysophanol has a half life of about 2.75hours.

Emodin

Also known as Rheum emodi, emodin has a molecular weight around 270.24g/mol. As to its anti-cancer activities, emodin exhibits proapoptoticity in prostate cancer cells through P53, and P21. Emodinincreases ROS, and improves chemotherapy effect in prostate cells whichare drug resistant. Emodin exhibits pro apoptoticity through inhibitionof IL6 in myltiple myaloma. Emodin exhibits anti-matastaticity0 throughintegrins effect. Emodin decreases HER2 in breast cancer and improveschemotherapoitic effect. Emodin induces cytotoxis in tongue carcinoma,and inhibits NFkB and other pro inflammatory cytokines. As to itsanti-inflammatory activities, emodin may be used as an anti ulcer agentthrough Hpylori destruction and change in gastric fluid. Emodin inducesa stabilizing effect on atherosclerotic plaque in vessels, and improvesinsulin and glucose changes in type 2 diabetes. Emodin promotes antiplasmodium against malaria, and may be used as an immunosuppressive,pesticide, purgative, spasmolytic, styptic, vasoelaxant, and/orviricide. Emodin has a half-life of approximately 227 min and convertsto two active metabolites through P450 in the liver.

Rhein

Also know as cassic acid, Rhein has a molecular weight around 284.22g/mol. As to its anti-cancer activity, rhein acts as an antiproliferative in hepatic carcinoma, breast cancer, SCC of lungs, andcervical cancer. Rhein improves taxol effect in breast cancer, andinhibits nasopharengeal carcinoma and EGFR. Rhein serves as a sinergistwith mitomycin. Rhein exhibits cytotoxicity in tongue carcinoma. Rheinmay be used for anti angiogenesis. As to its anti-inflammatory activity,rhein exhibits anti fibroticity, and promotes anti proliferation ofhepatic cells through inhibition of TGFb1. Rhein may be used as an antioxidant. Rhein decreases IL1B, and IL18 proinflammatory cytokines Rheinis also anti bacterial, and may be used against staph. Aunts. Rhein alsoincreases sensitivity to ADH (drug). Rhein has a half-life of about 205min (approx). Rhein is hydrophobic, and may be combined with lysin inorder to be hydrophilic, as rhein lysinate.

Chrysophanein

Chrysophanein exhibits significant in-vitro cytotoxic activity in cancercell lines

Rhapontin

Rhapontin has a molecular weight of around 420.41 g/mol. As to itsanti-cancer activities, rhapontin induces apoptosis and suppresses KATOIII cell-growth in stomach cancer. Rhapontin also provides protectiveeffects on LDL and erythrocytes against oxidative damage. Rhapontin hasa half life of about 23.5 minutes.

Stilbene Piceid

Stilbene piceid has a molecular weight around 390.2 g/mol. As to itsanti-cancer activities, stilbene piceid inhibits DNA synthesis in LLCcells. Stilbene piceid also has an inhibitory effect on lipoxygenase.Stilbene also exhibits antioxidant activity and inhibitsalpha-glucosidase. Stilbene piceid also inhibits lipid peroxidationinduced by ADP and NADPH in liver microsomes. Stilbene acts directly onsmooth muscle to promote pulmonary artery relaxation.

Tannin

Tannin has a molecular weight around 500-3000 g/mol. As to itsanti-cancer activity, tannin suppresses the growth of MCF-7 breastcancer cells. Geraniin, a form of tannin separated from geranium, causescell death through induction of apoptosis. Tannin exhibits differentantiproliferative effects against cervical and colon cancer cells grownin vitro. In pomegranate, tannin modulates inflammatory cell signalingin colon cancer cells. Tannin promotes apoptosis through induction ofp53 non-small cell lung cancer cells. As to its anti-inflammatoryactivity, tannin in tomato suppresses COX-2 expression. Tannin can beused as an in vitro antioxidant and/or antiplatelet and also as ananti-inflammatory due to its free radical scavenging effects. Tannin maybe used for its antiviral, antibacterial and/or antiparasitic effects.Tannin can be used in the treatment of HFE hereditary hemochromatosis.Tannin is capable of reversing 6-hydroxydopamine induced toxicity.Tannin has a dental use, as well, as tannin-fluoride preparation reducesgingival inflammation around abutment teeth. A large intake of tanninsmay cause bowel irritation, kidney irritation, liver damage, irritationof the stomach and/or gastrointestinal pain. A correlation has been madebetween esophogeal or nasal cancer in humans and regular consumption ofcertain herbs with high tannin concentrations. Tannins inhibit theabsorption of minerals such as iron which may, if prolonged, lead toanemia. Tannins are present in soil, plants, water, tea, wine, andfruit. Tannin has a half life of about 3.15 hours.

Carvacrol

Carvacrol, also known as cymophenol, has a molecular weight around150.217 g/mol. As to its anti-cancer activities, carvacrol promotesanti-tumor effects on human metastatic breast cancer cells, includingMDA-MB 231. Carvacrol is a potent inhibitor of cell growth inHumanNon-Small Cell Lung cancer. Carvacrol also inhibits growth of myoblastcells even after activation of mutated N-ras oncogene. As to itsanti-inflammatory activities, carvacrol activates PPAR and suppressesCOX-2 inflammation. Carvacrol may be used for its antiproliferative andantiplatelet properties. Carvacrol is present in the essential oil ofOriganum vulgare, oil of thyme, oil obtained from pepperwort, and wildbergamot. Carvacrol inhibits the growth of several bacteria strains,e.g. Escherichia coli and Bacillus cereus, and in pseudomonasaeruginosa, carvacrol disrupts the bacteria membrane. Carvacrol may beused for its antioxidant activity. Carvacrol has a half life of about1.29 hours.

Eugenol

Eugenol has a molecular weight of about 164.2 g/mol. Eugenol inducesapoptosis in human colon cancer cells, and inhibits invasion andangiogenesis of gastric carcinogenesis induced by MNNG. Eugenol in honeysignificantly inhibits the growth of Ehrlich ascites carcinoma.Eugenol-related biphenyl (S)-6,6′-dibromo-dehydrodieugenol elicitsspecific antiproliferative activity on neuroectodermal tumour cellspartially triggering apoptosis. Eugenol causes melanoma growthsuppression through inhibition of E2F1 transcriptional activity. Eugenolmay also be used as an antiseptic. Eugenol also inhibits plateletaggregation induced by agonists, including collagen, ADP and calciumionophore. Eugenol may be extracted from certain essential oilsespecially from clove oil, nutmeg, cinnamon, basil and bay leaf. Eugenolis also used in perfumeries, flavorings, essential oils and in medicineas a local antiseptic and anesthetic. Eugenol may be used for itsantioxidative properties. Eugenol exhibits hepatotoxicity. An overdoseof eugenol may induce convulsions, diarrhea, nausea, unconsciousness,dizziness, and/or rapid heartbeat. Eugenol can be allergenic. Eugenolmay express carcinogenicity through oxidative DNA damage by itsmetabolite. Eugenol has a half-life of 1.975 hours, and under certainconditions, may have a half life up to 4 hours or even 18 hours.

Geraniol

Geraniol has a molecular weight of about 154.25 g/mol. As to itsanti-cancer activities, Geraniol promotes an antiproliferative mechanismin human pancreatic adenocarcinoma cells. Geraniol also promotesanti-proliferative and cell cycle regulatory effects in human breastcancer cells. Geraniol can cause a 2-fold reduction of thymidylatesynthase and thymidine kinase expression in colon cancer cells.Geraniol, as a component of plant essential oils, sensitizes human coloncancer cells to 5-fluorouracil treatment. Geraniol suppresses pancreatictumor growth without significantly affecting blood cholesterol levels.As to its anti-inflammatory activities, Geraniol diminishes the levelsof inflammatory markers induced by pamidronate stimuli in vitro and invivo. Geraniol also promotes inhibitory effects on nitric oxide andprostaglandin E₂ production in macrophages. Geraniol is a component ofrose oil, palmarosa oil, and citronella oil, and small quantities ofgeraniol are in geranium and lemon, has a rose-like odor and is commonlyused in perfumes. Geraniol can be used as effective plant-based mosquitorepellent. Geraniol is found in cigarettes. As to biologic use,ion-exchange iontophoresis combined with geraniol is a highly effectivetransdermal delivery system. Geraniol suppresses Candida cell growth inthe vagina and its local inflammation when combined with vaginalwashing. Gernaiol is also allergenic. Geraniol has a half life of about0.713 hours.

Luteolin

Luteolin has a molecular weight of 286.24 g/mol. As to its anti-canceractivity, luteolin, particularly in combination with standard anticancerdrugs such as cisplatin, serves as a HDAC inhibitor, e.g., for thetreatment of lung cancer. Luteolin promotes synergistic/additive growthinhibitory effects and may be effective in chemoprevention treatment ofhead and neck and lung cancers. Luteolin induces G1 arrest in humannasopharyngeal carcinoma cells. Luteolin not only protects DNA fromoxidative damage, but also increases repair activity in Caco-2 cells. Alow concentration of Luteolin has little toxic effect on cancer cells,but such low concentrations can sensitize chemotherapeutic drugs invarious cancer cell lines. Luteolin selectively inhibitschymotrypsin-like and trypsin-like proteasome catalytic activities intumor cells. Luteolin inhibits invasion of prostate cancer PC3 cellsthrough E-cadherin.

Luteolin is a PDE4 inhibitor and a general phosphodiesterase inhibitor,and an Interleukin 6 inhibitor. Luteolin inhibits inflammatory responseand improves insulin sensitivity in the endothelium. Luteolin preventsLPS-induced TNF-α expression in cardiac myocytes through inhibitingNF-κB signaling pathway. Luteolin inhibits myelin basic protein-inducedhuman mast cell activation and mast cell-dependent stimulation of JurkatT cells. Luteolin inhibits cyclooxygenase-2 expression and scavengesreactive oxygen species.

Luteolin is found in leaves, but it is also seen in celery, thyme,dandelion, rinds, barks, clover blossom and ragweed pollen. Luteolin isuseful in the prevention and treatment of skin photoaging. Luteolininhibits microglia and alters hippocampal-dependent spatial workingmemory. Luteolin enhances insulin sensitivity via activation of PPARγtranscriptional activity in adipocytes. Luteolin can induce nausea,vomiting and gastric hypersecretion. Luteolin has a half life of about1.2 hours.

Saponins

As to the anti-cancer activities of saponins with oleanolic acid,achyranthoside H methyl ester, a novel oleanolic acid saponin derivativefrom Achyranthes fauriei roots, induces apoptosis in human breast cancerMCF-7 and MDA-MB-453 cells via a caspase activation pathway. Saponionwith oleanolic acid exhibit insecticidal activity against the Mexicanbean beetle larvae (Epilachna varivestis).

Vanillic Acid

Vanillic acid has a molecular weight of 168.14672 g/mol. Vanillic acidsuppresses metastatic potential of human cancer cells through PI3Kinhibition and decreases angiogenesis in vivo. Vanillic acid enhancesthe activity of human lymphocyte proliferation and secretion ofIFN-gamma. Vanillic acid has a beneficial effect on DSS-inducedulcerative colitis, thereby manifesting its usefulness in the regulationof chronic intestinal inflammation. Phenolic compounds in mushroomLentinula edodes (shiitake) are hepatoprotective through theirsuppression of immune-mediated liver inflammation. Vanillic acid isfound in the root of Angelica sinensis, and in olive oil. Vanillic acidpromotes reduced cellular tyrosinase activity, DOPA oxidase and melanincontents, as well as down-regulated expressions of melanocortin-1receptor (MC1R), microphthalmia-associated transcription factor (MITF),tyrosinase, and tyrosinase-related proteins 2 (TRP-2) and TRP-1.Vanillic acid contributes to the prevention of the development ofdiabetic neuropathy by blocking the methylglyoxal-mediated intracellularglycation system. There exist an oxidized form of vanillin. Vanillicacis has a half life of about 10.552 hours.

α-Terpineol

The anti-cancer activities of α-terpineol, or alpha-terpineol, arepartly mediated through the suppression of NF-kappaB activation.α-terpineol exhibits antiproliferative effects on erythroleukemic K562cells. α-terpineol inhibits gene expression of the IL-6 receptor.α-terpineol suppresses fMLP-, LPS- and PMA-stimulated superoxideproduction. α-terpineol is found in cajuput oil, pine oil, andpetitgrain oil, and is a common ingredient in perfumes, cosmetics, andflavors and tea. α-terpineol demonstrates different degrees of growthinhibition against 15 different genera of oral bacteria. α-terpineol cancause postural hypotension in pine oil, and can cause eye irritation.There are three isomers, alpha-, beta-, and gamma-terpineol.Alpha-terpineol has a half life of about 1.245 hours.

Aucubin

Aucubin has a molecular weight of 346.32978 g/mol. 1. Antiproliferativeactivity is through cell cycle arrest and apoptosis in human non-smallcell lung cancer A549 cells 2. research-DNA damage induced bytopoisomerase I poisoning as one of the possible mechanisms bywhichaucubin have shown antitumoral activity 3. research-cytotoxicactivity against MCF7-breast adenocarcinoma, HeLa-cervix adenocarcinoma,A431-skin carcinoma of epithelial origin 4. can obstructH(2)O(2)-induced apoptosis by regulating of the expression of Bcl-2 andBax, as well as suppression of caspases cascade activation.

Aucubin enhance the activity of human lymphocyte proliferation andsecretion of IFN-gamma. Aucubin is found in the leaves of Aucubajaponica (Cornaceae), Eucommia ulmoides (Eucommiaceae), and Plantagoasiatic (Plantaginaceae). Aucubin protects against liver damage inducedby carbon tetrachloride or alpha-amanitin, particularly when dosedintra-peritoneally. Aucubin provides neuroprotection in primary diabeticencephalopathy. Aucubin treatment can lower blood glucose. Aucubin canproduce an increase in the level of lipid peroxidation and a decrease inactivities of antioxidant enzymes in liver and kidneys. Aucubin has ahalf life of about 42.5 minutes.

Digoxin

Digoxin, also known as digitalis, has a molecular weight of 780.938g/mol. Digoxin treatment can inhibit HIF-1alpha synthesis and blocktumor growth. Digoxin induces apoptosis in a human acute T-celllymphoblastic leukemia cell line. Digoxin can serve as a specificneuroblastoma growth inhibitor and an unspecific inhibitor ofangiogenesis.

Digoxin is widely used in the treatment of various heart conditions,namely atrial fibrillation, atrial flutter and sometimes heart failurethat generally cannot be controlled by other medication. Digoxinincreases myocardial contractility, such that the heart rate isdecreased, while blood pressure increases as stroke volume is increased,leading to increased tissue perfusion. Digoxin improves myocardialefficiency due to improved hemodynamics, and improves the ventricularfunction curve. Digoxin affects the kidney by increased renal blood flowand increased GFR. A mild diuretic effect is seen typically only inheart failure. Digoxin may cause AV junctional rhythm and ectopic beats(bigeminy) resulting in ventricular tachycardia and fibrillation.

Digoxin can induce loss of appetite, nausea, vomiting and diarrhea asthe gastrointestinal motility increases. Other common effects of Digoxinare blurred vision, visual disturbances (yellow-green halos and problemswith color perception), confusion, drowsiness, dizziness, insomnia,nightmares, agitation, and depression, as well as a higher acute senseof sensual activities. Less frequent adverse effects of digoxin(0.1%-1%) include: acute psychosis, delirium, amnesia, convulsions,shortened QRS complex, atrial or ventricular extrasystoles, paroxysmalatrial tachycardia with AV block, ventricular tachycardia orfibrillation, and heart block. Dangerous interactions can occur betweendigoxin and verapamil, amiodarone, erythromycin, and epinephrine. Theefficacy of chemotherapeutic agent substrates of Pgp may be stronglyreduced in patients taking digoxin. Digoxin treatment increases the riskof invasive breast cancer among postmenopausal women. Digoxin has ahalf-life around 36 hours.

Isoacetoside

Isoacetoside, as an extract from Tecoma stans, exhibits a cytotoxiceffect on human hepatocarcinoma cells (Hep-G2). Isoacetoside has a halflife of about 3.7-6.4 hours.

β-Sitosterol

β-sitosterol, or beta-sitosterol, has a molecular weight around 414.71g/mol. β-sitosterol may be used to treat prostatic carcinoma and breastcancer. β-sitosterol may have chemopreventive potential by virtue of itsradical quenching ability in vitro, with minimal toxicity to normalcells. β-sitosterol also attenuates beta-catenin and PCNA expression,making it a potential anticancer drug for colon carcinogenesis.β-sitosterol significantly inhibits the growth, and induces theapoptosis, of SGC-7901 human stomach cancer cells in vitro. The decreaseof the bcl-2/bax ratio and DNA damage may produce apoptosis induced bybeta-sitosterol in SGC-7901 human stomach cancer cells. β-sitosterolenhances tamoxifen effectiveness on breast cancer cells by affectingceramide metabolism. β-sitosterol has a proapoptotic effect that ismediated through the activation of ERK and the block of the PI3K/Aktsignal pathway in MCA-102 cells. Therefore, beta-sitosterol has a strongpotential as a therapeutic agent for preventing cancers such asfibrosarcoma. Beta-sitosterol is an effective apoptosis-promoting agentand that incorporation of more phytosterols in the diet may serve apreventive measure for breast cancer. An anti-microtubule characteristicof beta-sitosterol may contribute to the proliferation inhibition ofSiHa cells in cervical cancer. β-sitosterol activates the sphingomyelincycle and induces apoptosis in LNCaP human prostate cancer cells.β-sitosterol promotes anti-asthmatic actions that may be mediated byinhibiting the cellular responses and subsequent release/synthesis ofTh2 cytokines β-sitosterol may have therapeutic potential in allergicasthma.

β-sitosterol is found in Nigella sativa, pecans, Serenoa repens (sawpalmetto), avocados, Curcurbita pepo (pumpkin seed), Pygeum africanum,cashew fruit, rice bran, wheat germ, corn oils, soybeans, sea-buckthorn,wolfberries, and Wrightia tinctoria.

β-sitosterol reduces blood levels of cholesterol, and is sometimes usedin treating hypercholesterolemia. β-sitosterol may produce a positiveeffect on male hair loss in combination with Saw palmetto. β-sitosterolmay play a major role in herbal therapy, especially in the treatment ofbenign prostatic hyperplasia. Beta-sitosterol is a naturally occurringphytosterol that may be used to cure atherosclerosis, diabetes, cancer,and inflammation and is also an antioxidant. β-sitosterol has a halflife of about 0.966 hours.

By taking more than the recommended dose of β-sitosterol, people maysuffer from stomach upset, nausea, diarrhea, gas or constipation,impotence (also known as erectile dysfunction or ED), decreased sexdrive. Beta-Sitosterol should be avoided during pregnancy andbreast-feeding, since it is not proven to be benign with regard topotential effects on unborn and newborn children. β-Sitosterol is alsonot recommended for individuals with sitosterolemia, a rare inheritedfat storage disease, because people with this condition have too muchβ-sitosterol and related fats in their system, taking β-sitosterol willonly worsen this condition. High levels of β-sitosterol concentrationsin blood have been correlated with increased severity of heart diseasein men having previously suffered from heart attacks, and may causeallergy.

Half lives have been indicated for certain molecules. The half lives ofmolecules can vary from these, e.g. generally based on the herb growingand/or preparation conditions, how it is combined with other herbs ofmolecules in treatment, or based on patient characteristics andbehaviors such as eating and drinking and physical activity, or onpotency or other factors. Doses and dose periods may be determined basedin part on the half lives. Typically, however, doses and dose periodswill be determined based on characteristics of the patient, thepatient's condition and the patient's history, as well as on theexpertise and experience of the attending physician.

The treatment method may include periodic doses including approximately5 ug/ml or 10 ug/ml or more of Emodin, along with respectively 0.05ug/ml or 0.10 ug/ml or more of digoxin. Other combinations may be usedin the treatment, including combining 5 ug/ml or more of emodin with atleast approximately 0.10 ug/ml digoxin, or at least approximately 10ug/ml emodin with at least approximately 0.10 ug/ml digoxin, or morethan 5 ug/ml of emodin with at least approximately 0.05 ug/ml digoxin,or at least approximately 10 ug/ml emodin and at least approximately0.05 ug/ml digoxin. Other combinations may be used and prescribed byphysicians depending on factors such variances in weight, age, gender,family or patient history, or other characteristics specific topatients.

The treatment regimen may include once or twice daily doses, or two ormore weekly doses weekly or otherwise. Doses may be taken more than onceor twice a day, while the amounts of each dose would be determinedaccording to the periodicity of the treatments.

Methods of preparing leukemia or other cancer treatment medicines arealso provided, including preparing a cocktail including a combination ofemodin and digoxin.

A leukemia or other cancer treatment medicine is also provided,including a cocktail including a combination of emodin and digoxin.

The present invention is directed to novel therapeutic applications ofcardiac glycosides such as digoxin and quabain alone or in combinationwith Emodin and its analogs for the treatment of cancer, and morespecifically, blood cancers. Unexpectedly, the present inventiondiscloses, for the first time, that digoxin synergize with emodin tostimulate cancer cell death.

FIG. 1 illustrates the effect of proliferation of digoxin, Emodin, andtheir combination on NB4 leukemic cells. The results represent averageof triplicates±STDEV. The percentage of life and death cells was doneusing staining for PI for death cells and the number of lived cells wasdetermined by FACS.

FIG. 2 illustrates the effect of proliferation of digoxin, Emodin, andtheir combination on HL-60 leukemic cells. The results represent averageof triplicates±STDEV. The percentage of life and death cells was doneusing staining for PI for death cells and the number of lived cells wasdetermined by FACS.

FIG. 3 illustrates the effect of proliferation of digoxin, Emodin, andtheir combination on Jurkat T cell leukemic cells. The results representaverage of triplicates±STDEV. The percentage of life and death cells wasdone using staining for PI for death cells and the number of lived cellswas determined by FACS.

FIG. 4 illustrates the effect of proliferation of digoxin, Emodin, andtheir combination on HT29 colon cancer cells. The results representaverage of triplicates±STDEV. The percentage of life and death cells wasdone using staining for PI for death cells and the number of lived cellswas determined by FACS.

FIG. 5 illustrates the effect of proliferation of digoxin, Emodin, andtheir combination on PC3 prostate tumor cells. The results representaverage of triplicates±STDEV. The percentage of life and death cells wasdone using staining for PI for death cells and the number of lived cellswas determined by FACS.

FIG. 6 illustrates the effect of proliferation of digoxin, Emodin andtheir combination on MDA 435 are cell line of Breast Cancer. In the invitro experiments, there is a dependency on concentration.

FIG. 7 illustrates the effect of proliferation of digoxin, Emodin andtheir combination on MDA 435 are cell lines of Melanoma and BreastCancer. In this case, concentration of 1/100 had no effect (same asControl) ⅓ had good effect and 1/10 killed all cells. Normal cells wereaffected 50-90% less than the rate of pathological cells. Treatmentsdescribed herein may be prepared for topical use for treatment ofmelanoma, BCC (basal cyr carcinoma) and inflammatory skin diseases likePsoriasis. For example, combinations of herbs and/or herbal extracts asdescribed herein may be prepared as a cream to apply onto the skin. Herbor herb extract combinations described herein also may be injected to toinfected areas of the skin of a patient using a syringe. An examplemethod for preparation of an external cream in accordance with certainembodiments is provided below.

Preperation of External Cream/Ointment

First, two of more of the herbs are cooked, for example, as describedelsewhere herein or as may be understood or determined by those skilledin the art. A cream is then prepared that may be somewhat more of lessthan half herbs and half cream, e.g., a 30% liquid of herbs in 1:1 ratioand 70% cream may be used. The herbs can also be prepared as a tincture,e.g., soaking the herbs in alcohol for a period of time such as 2 weeksin a ratio of 1:3, for example. This herbal liquid can then be mixedwith the cream in the same way as described above.

Treatments described herein may also be effective againstimmunodeficiency diseases such as HIV and AIDS, as well as otherconditions affecting the immune system.

FIG. 8 illustrates the effect of proliferation of digoxin, Emodin andtheir combination on Heparanase activity. Heparanase is an enzyme thatrelates to angiogenesis (the physiological process involving the growthof new blood vessels from pre-existing vessels). This slide depicts thefact that the proliferation of the combination of digoxin and Emodindelays the activity of this enzyme.

FIG. 9 illustrates the effect of proliferation of digoxin, Emodin andtheir combination on HMVEC which is also an enzyme that is related toangiogenesis.

FIGS. 10 and 11 illustrate the effect of proliferation of digoxin,Emodin and their combination on NB-4 is Cell Line for AML (Acute MiloLeukemia). The two left bars in the diagram of FIG. 10 are Control (DMSOas well). The Digoxin alone 0.1 ug/ml (illustrated by the 3rd bar) showsa strong effect. Emodin alone has an effect only in large concentration(approximately 10 ug/ml or more). The three bars to the right in FIG. 10are the effect of the combination of Digoxin and Emodin. Taking Digoxin0.1 ug/ml and Emodin 10 ug/ml has a very strong effect better thaneither of the two acting alone. In fact, the effect is more than doublewith 892 live cells measured when Emodin alone is used, and 892 whenDigoxin alone is used, and 371 were measured when both Digoxin andEmodin were used in combination. This surprising effect of thecombination of Digoxin and Emodin shows an advantageous feature of thepresent invention.

FIGS. 12-13 show that for HL-60, which is Promyolitic Leukemia, as wellas for Jurkat, which is Acute T cell Leukemia, the same surprisinglyadvantageous synergistic effect was observed.

FIG. 14 also shows a synergistic advantage of using the combination ofDigoxin and Emodin on the HT-29 cell line.

In certain embodiments, emodin and/or digoxin may be combined with oneor more of the other molecules described herein. In certain embodiments,any two or more of Emodin, Rhein, Rhapontin, Carvacrol, Vanillic acid,Sitosterol, Aucubin, Digoxin, and beta-sitosterol may be combined toform an effective treatment of diseases such as cancer, inflammatorydisease or auto immune deficiency. One or more further moleculesdescribed herein or otherwise may be combined with any one or more ofthese molecules as well.

EXAMPLES

The embodiments of the present invention are directed to advantageousmedicines and methods of treatment and preparation of medicines andtreatments for cancer wherein combinations of certain herbs such as DaHuang and Sheng Di Huang, and/or Jin Yin Hua, and/or Mu Dan Pi, Di GuPi, Xian He Cao and/or Chun Gen Pi, and/or any one or more of the elevenadditional herbs including Zi Cao, or radix arnebiae (arnebia root) orradix lithospermi (gromwell root), Xuan Shen, or radix scrophulariae(figwort root), Shi Gao or gypsum fibrosum (gypsum), Bai Shao, or radixpaeoniae alba (white peony root), Chi Shao or radix paeoniae rubra (redpeony root), Hong Hua or flos carthami (safflower), Da Qing Ye or foliumisatidis (woad leaf), Qing Dai or indigo naturalis (natural indigo), BaiZhu or rhizoma atractylodis macrocephalae (largehead atractylodesrhizome), Shi Wei or folium pyrrosiae (shearer's pyrrosia leaf), and/orRou Gui or cortex cinnamomi (cinnamomum bark), and/or combinations ofcertain herbal ingredients, such as digoxin “D” and emodin “E,” and/orone or more others described above including Emodin, Rhein, and/orRhapontin of Da Huang, Carvacrol, Vanillic acid, and/or Sitosterol ofJin Yin Hua, and/or Aucubin, Digoxin, and/or beta-sitosterol of Sheng DiHuang and/or other ingredients present within the two, the three, theseven and/or even the eighteen herbs, are used to inhibit tumor cellgrowth and/or reduce white cell count.

In one embodiment, doses of between 13.3 grams to 60 grams of Sheng DiHuang, and 3.3 grams to 30 grams of Da Huang are administered daily or2-3 times daily before meals or up to 10× daily or less than daily evenas little as three times a month. In other embodiments, the regimen mustdescribed further includes 3.3 grams to 30 grams of Jin Yin Hua. Inother embodiments, two of these three are included in the regimen. Infurther embodiments, one, two, three or all four of the herbs Mu Dan Pi,Xian He Cao, Chun Gen Pi and Di Gu Pi is/are combined with one, two orall of the three herbs Sheng Di Huang, Da Huang and Jin Yin Hua,including 3.3 grams to 15 grams of any of the other four of the sevenherbs, e.g., 3.3 grams to 15 grams of Mu Dan Pi, 3.3-10 grams to 15grams of Xian He Cao, 3.3-10 grams to 15 grams of Chun Gen Pi, and/or3.3-5 grams to 15 grams of Di Gu Pi.

In one example involving a combination of the three herbs, 40 grams ofSheng Di Huang in 100 grams of water, 15 grams of Da Huang in 60 gramsof water and 15 grams of Jin Yin Hua in 60 grams of water may becombined in a single treatement dosage and administered to a patent. Thecombination may be cooked once or twice, and the herb:liquid ratio maybe 1:10 or otherwise. The dose may be 1-10 days or even multiple timesdaily including 2-6 times daily, and even as often as 10 times daily.

In another example, a treatment regiment includes a combination of theseven herbs, such as 5 grams of Mu Dan Pi in 15 grams of water, 20 gramsof Sheng Di Huang in 80 grams of water, 5 grams of Xian He Cao in 15grams of water, 5 grams of Chun Gen Pi in 15 grams of water, 5 grams ofDi Gu Pi in 15 grams of water, 10 grams of Da Huang in 40 grams of waterand 10 grams of Jin Yin Hua in 40 grams of water. The combination may becooked once or twice, and the herb:liquid ratio may be 1:10 orotherwise. The dose may be 1-10 days or even multiple times dailyincluding 2-6 times daily, and even as often as 10 times daily.

In another example, a low dose formula involving the eighteen herbsincludes 3.3 grams of Da Huang in about 32 grams water, 13.3 grams ofSheng Di Huang in about 129 grams of water, 3.3 grams of Jin Yin Hua inabout 32 grams of water, 3.3 grams of Mu Dan Pi in about 32 grams ofwater, 5 grams of Di Gu Pi in about 48 grams of water, 10 grams of XianHe Cao in about 97 grams of water, 10 grams of Chun Gen Pi in about 97grams of water, 8.3 grams of Zi Cao in about 80 grams of water, 6.7grams of Xuan Shen in about 65 grams of water, 3.3 grams of Shi Gao inabout 32 grams of water, 4 grams of Bai Shao in about 39 grams of water,4 grams of Chi Shao in about 39 grams of water, 3.3 grams of Hong Hua inabout 32 grams of water, 8.3 grams of Da Qing Ye in about 80 grams ofwater, 6.7 grams of Qing Dai in about 65 grams of water, 6.7 grams ofBai Zhu in about 65 grams of water, 5 grams of Shi Wei in about 48 gramsof water, and 2 grams of Rou Gui in about 19 grams of water.

In another example, a high dose formula involving the eighteen herbsincludes 30 grams of Da Huang in about 300 grams water, 60 grams ofSheng Di Huang in about 600 grams of water, 30 grams of Jin Yin Hua inabout 300 grams of water, 15 grams of Mu Dan Pi in about 150 grams ofwater, 15 grams of Di Gu Pi in about 150 grams of water, 15 grams ofXian He Cao in about 150 grams of water, 15 grams of Chun Gen Pi inabout 150 grams of water, 5 grams of Zi Cao in about 50 grams of water,5 grams of Xuan Shen in about 50 grams of water, 5 grams of Shi Gao inabout 50 grams of water, 5 grams of Bai Shao in about 50 grams of water,5 grams of Chi Shao in about 50 grams of water, 5 grams of Hong Hua inabout 50 grams of water, 5 grams of Da Qing Ye in about 50 grams ofwater, 5 grams of Qing Dai in about 50 grams of water, 5 grams of BaiZhu in about 50 grams of water, 5 grams of Shi Wei in about 50 grams ofwater, and 5 grams of Rou Gui in about 50 grams of water.

In another example, a low dose formula involving the seven herbsincludes 3.3 grams of Da Huang in about 32 grams water, 13.3 grams ofSheng Di Huang in about 129 grams of water, 3.3 grams of Jin Yin Hua inabout 32 grams of water, 3.3 grams of Mu Dan Pi in about 32 grams ofwater, 5 grams of Di Gu Pi in about 48 grams of water, 10 grams of XianHe Cao in about 97 grams of water, and 10 grams of Chun Gen Pi in about97 grams of water.

In another example, a high dose formula involving the seven herbsincludes 30 grams of Da Huang in about 300 grams water, 60 grams ofSheng Di Huang in about 600 grams of water, 30 grams of Jin Yin Hua inabout 300 grams of water, 15 grams of Mu Dan Pi in about 150 grams ofwater, 15 grams of Di Gu Pi in about 150 grams of water, 15 grams ofXian He Cao in about 150 grams of water, and 15 grams of Chun Gen Pi inabout 150 grams of water.

In another example, a low dose formula involving the three herbsincludes 3.3 grams of Da Huang in about 32 grams water, 13.3 grams ofSheng Di Huang in about 129 grams of water, and 3.3 grams of Jin Yin Huain about 32 grams of water.

In another example, a high dose formula involving the three herbsincludes 30 grams of Da Huang in about 300 grams water, 60 grams ofSheng Di Huang in about 600 grams of water, and 30 grams of Jin Yin Huain about 300 grams of water.

In another example, a medium dose formula involving the three herbsincludes 10 grams of Da Huang in about 100 grams water, 25 grams ofSheng Di Huang in about 250 grams of water, and 10 grams of Jin Yin Huain about 100 grams of water.

In another example, a medium dose formula involving the three herbsincludes 20 grams of Da Huang in about 200 grams water, 40 grams ofSheng Di Huang in about 400 grams of water, and 20 grams of Jin Yin Huain about 200 grams of water.

In another example, a medium dose formula involving the three herbsincludes 15 grams of Da Huang in about 150 grams water, 35 grams ofSheng Di Huang in about 350 grams of water, and 15 grams of Jin Yin Huain about 150 grams of water.

In another example, a medium dose formula involving the three herbsincludes 25 grams of Da Huang in about 250 grams water, 45 grams ofSheng Di Huang in about 450 grams of water, and 25 grams of Jin Yin Huain about 250 grams of water.

In another example, a low dose formula involving the two herbs includes3.3 grams of Da Huang in about 32 grams water, and 13.3 grams of ShengDi Huang in about 129 grams of water.

In another example, a high dose formula involving the two herbs includes30 grams of Da Huang in about 300 grams water, and 60 grams of Sheng DiHuang in about 600 grams of water.

In another example, a medium dose formula involving the two herbsincludes 10 grams of Da Huang in about 100 grams water, and 30 grams ofSheng Di Huang in about 300 grams of water.

In another example, a medium dose formula involving the two herbsincludes 20 grams of Da Huang in about 300 grams water, and 45 grams ofSheng Di Huang in about 450 grams of water.

In another example, a medium dose formula involving the two herbsincludes 15 grams of Da Huang in about 32 grams water, and 40 grams ofSheng Di Huang in about 400 grams of water.

In another example, a medium dose formula involving the two herbsincludes 25 grams of Da Huang in about 300 grams water, and 50 grams ofSheng Di Huang in about 500 grams of water.

Examples of medium dose formulas for the seven herb and eighteen herbcombinations are also provided commensurate with the medium doseformulas for the two and three herbs combinations includingcorresponding percentage changes in the other four and other 15 herbs.Many other combinations may be used depending on characteristics of thepatient such as age and weight, the condition of the patient, and thepatient's history.

Doses in between the low dose and high dose examples for the two herbformula, the three herb formula, the seven herb formula, and theeighteen herb formula are also within the scope of further examples.Under certain conditions, doses above the high dose formula or below thelow dose formula may be used as determined by a physician using his orher expertise and experience both generally in the field and withspecific patients. In addition, other combinations of two, three, four,five, or more of these 18 herbs may be used in further examples offormulas wherein the dose ranges described in the above examples orotherwise as determined by a physician may be used. Further herbs notdescribed herein may also be included in formulas including combinationsof the described herbs.

In the examples above, certain doses of herb combinations have beendescribed in certain amounts of water. Mixing the herbs in water is onlyone example of a way to take the herbal combinations. Instead, no watermay be used and instead another liquid may be used. The herbs may beformed into pills or capsules, or an IV may be used. Any of the herbalcombinations may be formed into a cream and rubbed onto the skin, or asyringe may be used to inject a patient with an herbal combination.

In another example, the effect of CXCR4 antagonists on the survival ofNB4, HL60, Jurkat leukemic cells and HT29 colon cancer and prostatetumor cells was examined. Digoxin significantly inhibited the growth ofleukemic cells at concentrations between (0.05- to 1 microgram/ml).Emodin by itself inhibited the growth of leukemic cells only atconcentrations of more than 5 microgrm/ml. Combinations of digoxin atconcentrations of 0.1 microgram/ml and either 5 or 10 microgram/ml ofemodin increased significantly the tumor killing ability of bothcompounds. Digoxin, Emodin, and their combination also add partialhowever significant effect on HT29 tumor cells.

The effect of the proliferation of digoxin, Emodin, and theircombination on the viability of cancer cell lines from different originwas studied. Harvested non adherent human hematopoietic cancer celllines NB4, HL60 and Jurkat, cells were seeded at 2×105 viable cells/1 mlper well into a 24-well plate in triplicates in a medium supplementedwith 10% FCS and incubated with different concentrations of digoxin,Emodin, and their combination for 24 hours. Following the incubation,the cells were stained with propidium iodide (PI) (Sigma, St. Louis,Mo.) and percent of viable PI-negative cells in culture was determinedby FACScalibur analysis (Becton Dickinson Immunocytometry Systems),using CellQuest software. Adherent prostate cancer PC3 cells and coloncancer HT29 cells were seeded at 1×105 viable cells/1 ml per well into a24-well plate under conditions described above, and following a 24-hourexposure to digoxin, Emodin, and their combination the cells wereharvested, washed with PBS and stained with PI and counted as describedfor hematopoietic cells.

The therapeutic index of digoxin is understood to be between 0.125 m″gto 0.5 m″g, while the LD50 values of emodin in mice are understood to be0.56 g/k″g.

When combining D and E in accordance with the present invention, thereis a surprising and unexpected symbiotic effect.

The current treatment may be prepared as a mixture of herbs that areknown to have separately digoxin “D” and emodin “E”. Aloe-emodin is anatural anthraquinone compound that is present in some traditionalmedicinal plants such as Rhei Rhizoma and Rheum palmatum. Interestingly,aloe-emodin has been found to have lesser cytotoxicity towards normalhuman cells. In-vitro tests with synthesized D&E molecules have beenperformed on a cell-line. The in-vitro with the plants was similar tothe one with the isolated molecules.

Emodin may be extracted from traditional medicinal plants such as RheiRhizoma and Rheum Palmatum. In one embodiment, the source of Emodin usedis Da Huang—Chinese name, or Rhubarb Root—English name, or RheumPlamatum—Botanical name, or Radix Rhisoma Rhei-Pharmaceutical name.Emodin may be extracted from Rhubarb, Buckthorn and/or Japanese Knotweed(Fallopia Japonica). Aloe-emodin may be used which is a variety ofemodin found in Socotrine, Barbados, and Zanzibar aloes.

A medicinal cancer treatment using Emodin is prepared in certainembodiments by mixing the herb, Da Huang, in water at a ratio ofapproximately 10:1. The herb may be ground to a fine powder. The watermay be added to the fine powdered herb, and the pot covered. Afterboiling, the heat is lowered in certain embodiments to about 70 degreescentigrade. The aqueous mixture is cooked for another hour. The liquidis then strained into a container. In some cases, this may be donetwice. In the second cooking, the ratio may be reduced to 7.5:1. Thesecond cooking may take about 45 minutes including the boiling.

The source of Digoxin in certain embodiments is Sheng Di Huang-Chinesename, or Foxglove root—English name, or Radix Rhemania—Pharmaceuticalname. The preparation of the Digoxin may be the same as for the Emodin.In certain embodiments, the herbs from which the Emodin and Digoxin areextracted are cooked together. Digoxin may be extracted from DigitalisPurpurea or Purple Foxglove.

In certain embodiments, the preparede treatment is not pure. Forexample, certain treatments may involve a “vegetable soup” type regimentmade of more than one and even two, three, seven or eighteen herbalingredients per specific examples provided herein, or any combinationsof the herbs described herein, or combinations of the herbs describedherein with other herbs not mentioned herein. In certain embodiments,herbal ingredients are mixed in solution and a patient may drink theliquid. While freeze-drying the herb in powder form may be possible, theabove-described process appears to be more effective.

Harvested non adherent human hematopoietic cancer cell lines NB4 (AML),HL60 and Jurkat were seeded at 2×105 viable cells/1 ml per well into a24-well plate in triplicates in a medium supplemented with 10% FCS andincubated with different concentrations of digoxin, Emodin, and theircombination for 24 hours. Following the incubation, the cells werestained with propidium iodide (PI) (Sigma, St. Louis, Mo.) and percentof viable PI-negative cells in culture was determined by FACScaliburanalysis (Becton Dickinson Immunocytometry Systems), using CellQuestsoftware. Adherent prostate cancer PC3 cells and colon cancer HT29 cellswere seeded at 1×105 viable cells/1 ml per well into a 24-well plateunder conditions described above, and following 24-hour exposure todigoxin, Emodin, and their combination, the cells were harvested, washedwith PBS and stained with PI and counted as described for hematopoieticcells.

INCORPORATION BY REFERENCE

What follows is a cite list of references which are, in addition tothose references cited above and below herein, and including that whichis described as background, the invention summary, brief description ofthe drawings, the drawings and the abstract, hereby incorporated byreference into the detailed description of the preferred embodimentsbelow, as disclosing alternative embodiments of elements or features ofthe preferred embodiments not otherwise set forth in detail below. Asingle one or a combination of two or more of these references may beconsulted to obtain a variation of the preferred embodiments describedin the detailed description below. Further patent, patent applicationand non-patent references are cited in the written description and arealso incorporated by reference into the preferred embodiments.

The cancer treatment including administering a cocktail including acombination of emodin and digoxin may be combined with other treatmentssuch as may be understood by those skilled in the art and/or as may bedescribed in literature such as the following which are herebyincorporated by reference as disclosing alternative embodiments andcompounds that may be combined with an emodin, digoxin cocktail in aneffective cancer therapy:

-   Chronic Lymphocytic Leukemia by the Leukemia & Lymphoma Society,    Medifocus.com Medifocus Guide on Chronic Lymphocytic Leukemia;-   RANJIT THOMAS, et al., Spontaneous Clinical Regression in Chronic    Lymphocytic Leukemia, British Journal of Haematology, 2002, 116,    341-345;-   DRAGOMIR MARISAVLJEVIC, et al., Spontaneous Clinical Remission of    Chronic Lymphocytic Leukemia, Haema 2003; 6(3): 394-397;-   UPSHAW J D Jr., et al., Spontaneous Remission of B cell Chronic    Lymphocytic Leukemia associated with T Lymphocytic Hyperplasia in    bone marrow, South Med J. 22 Jun. 1995(6): 647-9;-   WIERNIK P H, Second neoplasms in patients with chronic lymphocytic    leukemia, Current Treat Options Oncol. 2004 June; 5(3):215-23;-   LUIS FAYAD M D and SUSAN O'BRIEN M D, Chronic Lymphocytic Leukemia    and Associated Disorders, Medical Oncology: A Comprehensive Review,    1995;-   MICHAEL J. KEATING, et al., Biology and Treatment of Chronic    Lymphocytic Leukemia, American Society of Hematology, Hematology    2003, 153-175;-   G E MARTI, et al, Descriptive Epidemiology of Chronic Lymphocytic    Leukemia (CLL); and U.S. Pat. Nos. 7,790,905, 7,695,926, 7,563,584,    7,393,656, 7,381,535, 7,358,222, 7,268,162, 6,812,255, 6,740,665,    6,197,754, 5,872,103 and published patent applications serial nos.    WO2006053049, US20050196473, US20030211180, WO0166123, WO2007130124,    WO2004052294, US20100092585, US20100167286, US20100144647,    US20100068198, US20090269772, US20090215042, US20090143279,    US20080152700, US20080220441, US20070191262, US20060205679, and    US20050026849.

It is also believed that treatment with an advantageous combination ofEmodin and Digoxin may be provided to treat other ailments such aspsoriasis and inflammatory disease.

It is contemplated, as people with ordinary skill in the art would do,that the newly separated compounds may be each individually or incombination used as an ingredient to prepare a pharmaceuticalcomposition for a particular treatment purpose. As it is the status ofthe art in the pharmaceutical industry, once substantially purepreparations of a compound are obtained, various pharmaceuticalcompositions or formulations can be prepared from the substantially purecompound using conventional processes or future developed processes inthe industry. Specific processes of making pharmaceutical formulationsand dosage forms (including, but not limited to, tablet, capsule,injection, syrup) from chemical compounds are not part of the inventionand people of ordinary skill in the art of the pharmaceutical industryare capable of applying one or more processes established in theindustry to the practice of the present invention. Alternatively, peopleof ordinary skill in the art may modify the existing conventionalprocesses to better suit the compounds of the present invention. Forexample, the patent or patent application databases provided at USPTOofficial website contain rich resources concerning making pharmaceuticalformulations and products from effective chemical compounds. Anotheruseful source of information is Handbook of Pharmaceutical ManufacturingFormulations, edited by Sarfaraz K. Niazi and sold by Culinary &Hospitality Industry Publications Services, which is incorporated byreference.

While the invention has been described in terms of several embodiments,those skilled in the art will recognize that the invention is notlimited to the embodiments described, but can be practiced withmodification and alteration within the spirit and scope of the appendedclaims. The description is thus to be regarded as illustrative insteadof limiting of the invention as set forth in the appended claimsincluding structural and functional equivalents thereof.

What is claimed is:
 1. A method of preparing a medicinal combination fortreating a patient diagnosed with leukemia, melanoma, psoriasis orbreast, prostate or colon cancer, comprising preparing a combination ofeffective amounts of each of Da Huang, Sheng Di Huang and Jin Yin Hua,wherein the Sheng Di Huang, the Da Huang and the Jin Yin Hua togethercomprise 57% or more of said combination.
 2. The method of claim 1,wherein the Sheng Di Huang comprises 33% or more of said combination. 3.The method of claim 1, comprising 3.3 grams or more of Da Huang and 13.3grams or more of Sheng Di Huang and 3.3 grams or more of Jin Yin Hua. 4.A method of treating a patient diagnosed with leukemia, melanoma,psoriasis or breast, prostate or colon cancer, comprising administeringto said patient a combination of effective amounts of each of Da Huang,Sheng Di Huang and Jin Yin Hua, wherein the Sheng Di Huang, the Da Huangand the Jin Yin Hua together comprise 57% or more of said combination.5. The method of claim 4, wherein the Sheng Di Huang comprises 33% ormore of said combination.
 6. The method of claim 4, wherein thecombination comprises 3.3 grams or more of Da Huang and 13.3 grams ormore of Sheng Di Huang and 3.3 grams or more of Jin Yin Hua.
 7. Amedicinal combination for treating a patient diagnosed with leukemia,melanoma, psoriasis or breast, prostate or colon cancer, comprisingeffective amounts of each of Da Huang, Sheng Di Huang and Jin Yin Hua,wherein the Sheng Di Huang, the Da Huang and the Jin Yin Hua togethercomprise 57% or more of said medicinal combination.
 8. The medicinalcombination of claim 7, wherein the Sheng Di Huang comprises 33% or moreof said medicinal combination.
 9. The medicinal combination of claim 7,comprising 3.3 grams or more of Da Huang and 13.3 grams or more of ShengDi Huang and 3.3 grams or more of Jin Yin Hua.